CN109430022A - The measuring method of ice plant water planting implantation methods and LED light matter proportion - Google Patents
The measuring method of ice plant water planting implantation methods and LED light matter proportion Download PDFInfo
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- CN109430022A CN109430022A CN201811171787.2A CN201811171787A CN109430022A CN 109430022 A CN109430022 A CN 109430022A CN 201811171787 A CN201811171787 A CN 201811171787A CN 109430022 A CN109430022 A CN 109430022A
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- ZLMJMSJWJFRBEC-UHFFFAOYSA-N potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
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- 229910052700 potassium Inorganic materials 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Abstract
The invention belongs to plant factor's water planting and cultivation technique without soil field, the measuring method of a kind of ice plant water planting implantation methods and LED light matter proportion, i.e. feux rouges: blue light: ultraviolet light=20:5:1 are disclosed；3 fluorescent tubes are installed, intensity of illumination is 300 μm of ol/ (m2S), period 14h/d, room temperature are 20-22 DEG C.Experiment carries out 3 repetitions, repeats that 7 kinds of light qualities are arranged every time, feux rouges, blue light, feux rouges: blue light=4:1, feux rouges: blue light=8:1, feux rouges: blue light: green light=4:1:1, feux rouges: blue light: ultraviolet light=20:5:1, white light control；Wherein the wavelength of feux rouges is (660 ± 10) nm, the wavelength of blue light is (450 ± 10) nm, the wavelength of green light is (520 ± 10) nm, the wavelength of ultraviolet light is (300 ± 10) nm.Control selects 18W white fluorescent lamp as light source.
Description
Technical field
The invention belongs to water planting and cultivation technique without soil field more particularly to a kind of ice plant water planting implantation methods
And the measuring method of LED light matter proportion.
Background technique
Currently, the prior art commonly used in the trade is such that illumination is one of the fundamental of plant growth and development,
Production and quality responses for crop, which have, to be significantly affected, light emitting diode (light-emittingdiode, abbreviation LED)
Novel semi-conductor light source, have many advantages, such as can bill coloured light, small in size, fever less, the service life is long, efficiency is high, become and ground
Study carefully the ideal tools of Illumination on Plant growth and development influence, LED light source plays in facilities horticulture research and production application at present
Important function.Light emitting diode can issue monochromatic light required for plant, and can need to incite somebody to action according to different plants
Monochromatic light independent assortment utilizes, and improves plant to the utilization efficiency of luminous energy.It is reported that by adjusting LED light in facility cultivation
Matter and light intensity can effectively improve the photosynthetic efficiency of the vegetables such as romaine lettuce, cucumber, shoot vegetable, promote vegetable growth, improve vegetables
Quality.Also rare report is studied in influence in relation to LED illumination to ice plant (being also ice dish) growth and development.Ice Ye Zhong
Flower originates in Africa, starts recent years in Chinese prevalence, but correlative study is also at the early-stage, and the people of research is also fewer, and
And LED and plant factor's technology are all the technologies that latest development is got up, therefore research is less, especially LED is to ice plant
Influence researches ice plant as a kind of novel rare characteristic nutritive health-care vegetable, nutritive value with higher
And economic value.Ice plant is a kind of Soiline-alkali plants, can grow at seashore, contain salt in the liquid of bulliform cell
Point, therefore taste inherently with some saline tastes.It is eaten in some places in Europe as vegetables, is also passed to the country now.Ice dish
Rich in the high substance of the functionals such as amino acid, anti-acid substance.Tart flavour in ice dish is the tart flavour of natural apple, and rich
Containing minerals such as sodium, potassium, carrotene, one of the characteristic of cabbage, Chinese cabbage, lettuce etc. has been agglomerated, has been the vegetable of high nutritive value
Dish.
In conclusion problem of the existing technology is: the influence research in relation to LED illumination to ice dish growth and development is also
Rare report, it is still necessary to explore for the LED planting technology of ice dish.
It solves the difficulty and meaning of above-mentioned technical problem: technical difficulty: being grown for measuring different light medium with ice dish is compared
And the determination and measurement and the measurement of Different Nutrition ingredient and analysis of nutritional quality influence measurement index；Meaning: for life
The quality of light filling processing enhancing ice dish has important references in the plantation of plant factor's high-quality and common plantation of ice dish in production
Meaning.
Summary of the invention
In view of the problems of the existing technology, the present invention provides a kind of ice plant water planting implantation methods and LED lights
The measuring method of matter proportion
The invention is realized in this way a kind of ice plant water planting implantation methods, the ice plant water planting plantation
Method includes: feux rouges: blue light: ultraviolet light=20:5:1；3 fluorescent tubes, photoperiod 14h/d, room temperature 20-22 are installed
℃。
Another object of the present invention is to provide a kind of ice plant water planting implantation methods, which is characterized in that the ice
Ye Zhong flower water planting implantation methods carry out nursery using nursery sponge, and sponge is saturating with water-wet, and seed is directly sowed in nursery sea
In silk floss, it is protected from light processing 2 days, seed is sprouted, and gives illumination later, when seedling grows to two leaves wholeheartedly, selects the consistent children of growing way
Seedling carries out water culture experiment；Nutrient solution uses Hogland nutrient solution, solid cultivation, and water planting frame is divided into 3 layers, and 1 layer can plant ice dish 36
, every layer is arranged a kind of light source；It is high every the growth indexes of survey in 10 days, including stem since after transplanting 15 days, leaf logarithm,
Blade is long, and blade is wide, and hat width is long and hat width is wide, and every plant of fresh weight and ground are collected and measured to plant by continuous measurement 4 times
Kind, and calculate root/shoot ratio；Collected material is subjected to Vc, chlorophyll a, b, the survey of total protein and total reducing sugar ingredient in laboratory
Amount, every layer takes 6 samples to be measured, and is averaged, 3 repetitions of every kind of light.
Another object of the present invention is to provide a kind of measurement sides of the LED light matter of ice plant water planting plantation
Method, the measuring method of the LED light matter of ice plant water planting plantation the following steps are included:
Step 1, experiment carry out 3 repetitions, repeat that 7 kinds of light qualities are arranged every time, feux rouges, blue light, feux rouges: blue light=4:1,
Feux rouges: blue light=8:1, feux rouges: blue light: green light=4:1:1, feux rouges: blue light: ultraviolet light=20:5:1, white light control；
Step 2, control select 18W white fluorescent lamp as light source；
3 fluorescent tubes, photoperiod 14h/d are installed in step 3, the top of every kind of processing, and room temperature is 20-22 DEG C.
Further, the Vc measuring method includes:
(1) Sample pretreatment takes tissue block 0.2g-0.5g that can at least rinse in ice-cold PBS to 5-10mg, filter
Paper wipe it is dry, weighing, be put into the homogenate tube of 5ml；
(2) the homogenate medium of 4 times of volumes is added in homogenate tube in g by weight: volume ml=1:4 ratio, ice-water bath item
Under part, tissue block is shredded as early as possible with small cut of ophthalmology；
(3) mode being homogenized: manual homogenization, left hand hold homogenate tube and fill lower end insertion in the vessel of mixture of ice and water,
Stamp stem is inserted perpendicularly into casing by the right hand, is rotated upwardly and downwardly grinding tens of times, is sufficiently ground, 20% homogenate is made.
(4) by the 20% homogenate generic centrifuge prepared or 4000 revs/min of low temperature low speed centrifuge, it is centrifuged 10-
15 minutes, supernatant is taken to be measured；
(5) prepared by supernatant: taking sample 0.15ml reagent adding one to apply liquid 0.45ml, vortex mixes, after placing 15 minutes
Centrifugation, is centrifuged 10 minutes by 3500-4000 revs/min, and upper layer clear liquid is supernatant.
Further, in the supernatant VC measurement: 37 DEG C mix well for water-bath 30 minutes, stand 10 minutes, wavelength
536nm, optical path 1cm, distilled water zeroing, measure each pipe absorbance value；
Calculation formula:
VC content (mcg/ml)=(measurement OD value-blank OD value)/(standard OD value-blank OD value) * standard items are dense
Spend (6 mcg/ml) * Sample pretreatment extension rate (4 times) * 4.
Further, the chlorophyll measuring method includes:
(1) fresh plant blade is taken, middle arteries are removed for plant leaf blade, 0.1g is weighed and shreds, distilled water cleans up；
(2) mixing well dehydrated alcohol and acetone according to 1:2v:v ratio will add for extracting solution in processing sample for use
Enter 1ml distilled water, pulvis 50mg is fully ground under the conditions of being protected from light and is placed in 10ml teat glass；
(3) mortar is rinsed with extracting solution, washing lotion is transferred to teat glass, extracting solution is settled to 10ml, is being protected from light condition
Lower extraction about 3h is placed in glass using masking foil if it cannot be protected from light and tries until observing that bottom residues bleach lower extraction completely
Shield lights on pipe；
(4) leaching liquor extracts 1ml and 1ml glass cuvette, and extracting solution zeroing measures extinction at 663 and 645nm respectively
Degree, respectively A663 and A645；
Calculation formula:
Chlorophyll-a Content (mg/g fresh weight)=(12.7 × A663-2.69 × A645) × V mentions (10ml) × extension rate
÷m/(g)÷1000；
Content of chlorophyll b (mg/g fresh weight)=(22.9 × A645-4.68 × A663) × V mentions (10ml) × extension rate
÷m/(g)÷1000；
Chlorophyll content (mg/g fresh weight)=(20.21 × A645+48.02 × A663) × V proposes (10ml) × dilution times
Number ÷ m/ (g) ÷ 1000.
Further, the method for protein measurement includes:
(1) Sample pretreatment takes tissue 0.2g-0.5g) it can at least be rinsed in ice-cold PBS to 5-10mg, filter paper
Wipe dry, correct amount is put into the homogenate tube of 5ml；
(2) the homogenate medium of 4 times of volumes is added in homogenate tube in g by weight: volume ml=1:4 ratio, ice-water bath item
Under part, tissue block is shredded as early as possible with small cut of ophthalmology；
(3) mode being homogenized: manual homogenization, left hand hold homogenate tube and fill lower end insertion in the vessel of mixture of ice and water,
Stamp stem is inserted perpendicularly into casing by the right hand, is rotated upwardly and downwardly grinding tens of times, is sufficiently ground, 20% homogenate is made；
(4) by the 20% homogenate generic centrifuge prepared or 4000 revs/min of low temperature low speed centrifuge, it is centrifuged 10-
15 minutes, supernatant is taken to be measured.
Further, the measuring method of the supernatant includes: to mix, and stands 10 minutes, wavelength 595nm, optical path 1cm, double
Water zeroing is steamed, each pipe absorbance value is measured；
Calculation formula:
Sample to be tested protein concentration (gprot/L)=" (measurement OD value-blank OD value)/(standard OD value-blank OD value) "
Extension rate before × standard concentration (0.563g/L) × sample measures.
Further, the plant soluble sugar measuring method includes: to weigh sample to be tested 0.1-0.2g, according to test sample
Distilled water is added in this quality g: distilled water ml=1:10 ratio, is ground into homogenate with homogenizer, pours into and change centrifuge tube
In, boiling water bath 10min；It covers tightly, pipe, which covers, pricks an aperture, and after cooling, 4000 revs/min, room temperature is centrifuged 10min, takes supernatant
It is diluted with 10 times of distilled water, shakes up that prepare sample supernatant spare；
Calculation formula:
Solvable sugared content (micro- gram gram of weight in wet base)=(measurement OD- blank OD)/(standard OD- blank OD) * standard concentration
Extension rate before (100 mcg/ml) ÷ sample to be tested fresh weight/(10 × homogenate distilled water volume ml) × sample measures.
In conclusion advantages of the present invention and good effect are as follows: the present invention from the point of view of the speed of growth and increment, 8:1 and
The speed of growth of 20:5:1 light processing is most fast, increment is maximum (Fig. 3), and fresh weight and on the ground weight are also maximum (Fig. 4).From life
From the point of view of long quality, either Vc content or soluble protein and soluble sugar content, 20:5:1 are superior to other light processings
(Fig. 6), in terms of Chlorophyll content and chlorophyll a and content of chlorophyll b, 20:5:1 also shows superiority, only with
Other processing ratios, difference is not significant (Fig. 5).The plant strain growth gesture and increment of white light control and blue light processing are minimum, followed by
Full feux rouges processing, it is seen then that red blue light matter exists simultaneously the growth for being more advantageous to ice dish, and comprehensively consider ice dish increment and
Nutritional quality, 20:5:1 are the light proportions most accommodated, it is seen that the addition of a small amount of UV-B is more advantageous to Vc, solubility in ice dish
The raising of sugar and soluble protein content improves the nutritional quality of ice dish.This result is high for the plant factor for producing upper ice dish
The quality of light filling processing enhancing ice dish has important references meaning in quality plantation and common plantation.
Detailed description of the invention
Fig. 1 is ice plant water planting implantation methods flow chart provided in an embodiment of the present invention.
Fig. 2 is the measuring method of the LED light matter proportion of ice plant water planting implantation methods provided in an embodiment of the present invention
Flow chart.
Fig. 3 is the growth curve of ice dish difference growth parameter(s) under different light medium proportion processing provided in an embodiment of the present invention
Schematic diagram.
Fig. 4 is the weighing results schematic diagram of ice dish under different light medium proportion processing provided in an embodiment of the present invention.
Fig. 5 is the chlorophyll content schematic diagram of ice dish under different light medium proportion processing provided in an embodiment of the present invention.
Fig. 6 is under different light medium proportion processing provided in an embodiment of the present invention, and the nutrition Quality Analysis result of ice dish is illustrated
Figure.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to this hair
It is bright to be further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, not
For limiting the present invention.
Application principle of the invention is explained in detail with reference to the accompanying drawing.
As shown in Figure 1, ice plant water planting implantation methods provided in an embodiment of the present invention include: feux rouges (R): blue light
(B): ultraviolet light (UV-B)=20:5:1；3 fluorescent tubes, photoperiod 14h/d are installed, room temperature is 20-22 DEG C.
S101: nursery is carried out using nursery sponge, sponge is saturating with water-wet, and seed is directly sowed in nursery sponge, is kept away
Light processing 2 days, seed was sprouted, and gave illumination later, when seedling grows to two leaves wholeheartedly, the consistent seedling of growing way was selected to carry out
Water culture experiment；Nutrient solution uses Hogland nutrient solution；
S102: solid cultivation, water planting frame are divided into 3 layers, and 1 layer can plant ice dish 36, and every layer is arranged a kind of light source；From transplanting
Start within 15 days afterwards, high every the growth indexes of survey in 10 days, including stem, leaf logarithm, blade is long, and blade is wide, hat width length and hat width
Width, continuous measurement 4 times；
S103: plant is collected and is measured every plant of fresh weight and is planted on the ground, and calculates root/shoot ratio；By collected material
Vc, chlorophyll a, b are carried out in laboratory, the measurement of total protein and total reducing sugar ingredient, every layer takes 6 samples to be measured, and is averaged
Value, 3 repetitions of every kind of light.
As shown in Fig. 2, the survey that the LED light matter of ice plant water planting implantation methods provided in an embodiment of the present invention matches
Determine method the following steps are included:
S201, experiment carry out 3 repetitions, repeat that 7 kinds of light qualities are arranged every time, feux rouges (R), blue light (B), feux rouges (R): blue
Light (B)==4:1, feux rouges (R): blue light (B)=8:1, feux rouges (R): blue light (B): green light (G)=4:1:1, feux rouges (R): blue
Light (B): ultraviolet light (UV-B)=20:5:1, white light control；
S202, control (CK) select 18W white fluorescent lamp (Ou Pu) to be used as light source；
3 fluorescent tubes, photoperiod 14h/d are installed in S203, the top of every kind of processing, and room temperature is 20-22 DEG C.
Application effect of the invention is described in detail below with reference to experiment.
Experimental material is Japan musashi open country cenospecies for examination ice plant seed, has purchased from Chongqing Jiang Yi seed sale
Limit company.It is provided for examination LED lamp tube by China Energy Saving Latticelighting Co., Ltd., every power of lamp tube is 9W.With ice plant
For test material, under the conditions of water planting, 7 kinds of light qualities are provided with, feux rouges, blue light, feux rouges: blue light=4:1, feux rouges: blue light=8:1, red
Light: blue light: green light=4:1:1 (like white light on sense organ) and feux rouges: blue light: ultraviolet light (UV-B)=20:5:1, white light control,
Measurement analyzes influence of the different light medium to ice plant growth and development and nutritional quality.
Experimental method, experiment carry out in plant factor, Jiangxi Prov. Academy of Science 2 months -2018 years in March, 2016, test
3 repetitions are carried out, repeat that 7 kinds of light qualities, feux rouges (R), blue light (B), feux rouges (R): blue light (B)==4:1, feux rouges is arranged every time
(R): blue light (B)=8:1, feux rouges (R): blue light (B): green light (G)=4:1:1, feux rouges (R): blue light (B): ultraviolet light (UV-B)
=20:5:1, white light control, wherein the wavelength of feux rouges is (660 ± 10) nm, the wavelength of blue light is (450 ± 10) nm, green light
Wavelength is (520 ± 10) nm, the wavelength of ultraviolet light is (300 ± 10) nm.It compares (CK) and selects 18W white fluorescent lamp (Ou Pu)
As light source.3 fluorescent tubes, photoperiod 14h/d are installed in the top of every kind of processing, and room temperature is 20-22 DEG C.
1, growth measurement
Nursery is carried out using nursery sponge in plant factor, sponge is saturating with water-wet, and seed is directly sowed in nursery sea
In silk floss, it is protected from light processing 2 days, seed is sprouted, and gives illumination later, when seedling grows to two leaves wholeheartedly, selects the consistent children of growing way
Seedling carries out water culture experiment.Nutrient solution uses Hogland nutrient solution, solid cultivation, and water planting frame is divided into 3 layers, and 1 layer can plant ice dish 36
, every layer is arranged a kind of light source.It is high every the growth indexes of survey in 10 days, including stem since after transplanting 15 days, leaf logarithm,
Blade is long, and blade is wide, and hat width is long and hat width is wide, continuously measures 4 times and (reaches highest increment, it is raw to start progress reproduction later
Long, plant type is smaller and smaller, and the leaf also the long smaller, starts to bloom by 60 days or so), plant is collected to and is measured every plant fresh
Weight and on the ground kind, and calculate its root/shoot ratio.Collected material is subjected to Vc, chlorophyll a, b in laboratory, total protein and total
The measurement of sugared ingredient, every layer takes 6 samples to be measured, and takes its average value, 3 repetitions of every kind of light.
2, VC measuring method
2.1 measuring principle
This law is acted on generated Fe2+ rapidly with Fe3+ and reduction type ascorbic acid, and the latter, can again with coffee sieve quinoline chromogenic reaction
To measure ascorbic content in blood plasma.
Reagent composition and one stock solution of reagent preparation
The application liquid of reagent one is prepared: the used time is according to one stock solution of reagent: distilled water=1:14 dilution mixes preparation.
Reagent two: acne
The application liquid of reagent two is prepared: one acne of used time add 0.36ml glacial acetic acid to add water to 40ml, dissolution, 4 DEG C of guarantors
It deposits.
Reagent three: white crystals acne
The application liquid of reagent three is prepared: because sufficiently dissolving system with first 2-4 hours plus 95% dehydrated alcohol 60ml compared with indissoluble solution
It is standby, 4 DEG C of preservations.
Yellow stock solution 0.2ml
The application liquid of reagent four is prepared: the used time takes four stock solution of 0.15ml reagent to add water to 25ml preparation, (matching while using).
Reagent five: liquid 6ml
Six: VC standard items of reagent: pulvis 6mg, every bottle includes vitamin C
The preparation of 600 mcg/ml standard items application liquid: 1 standard items pulvis is dissolved in 10ml reagent one using liquid
In, preparation.
The preparation of 6 mcg/ml VC standard items application liquid: 600 mg/ml VC standard items application liquid are taken to use reagent again
One applies 100 times of spare, matching while using of dilution of liquid.
2.2, operating procedure
(1) Sample pretreatment
Take tissue block (0.2g-0.5g) at least can be to 5-10mg in ice-cold PBS (PH7.4 phosphate buffer)
Rinsing, filter paper are wiped dry, and correct amount is put into the homogenate tube of 5ml.
(2) by weight (g): the homogenate medium of 4 times of volumes is added in homogenate tube in volume (ml)=1:4 ratio, ice water
Under the conditions of bath, tissue block is shredded as early as possible with small cut of ophthalmology.
(3) mode being homogenized: manual homogenization
Left hand holds homogenate tube and fills lower end insertion in the vessel of mixture of ice and water, and stamp stem is inserted perpendicularly into casing by the right hand
In, tens of times of grinding (6-8 minutes) is rotated upwardly and downwardly, sufficiently grinds, 20% homogenate is made.
(4) by the 20% homogenate generic centrifuge prepared or 4000 revs/min of low temperature low speed centrifuge, it is centrifuged 10-
15 minutes, supernatant is taken to be measured.
(5) prepared by supernatant: taking sample 0.15ml reagent adding one to apply liquid 0.45ml, vortex mixes, after placing 15 minutes
Centrifugation, is centrifuged 10 minutes by 3500-4000 revs/min, and upper layer clear liquid is supernatant.
The measurement of VC in supernatant:
It mixes well, 37 DEG C mix well for water-bath 30 minutes, stand 10 minutes, wavelength 536nm, optical path 1cm, distilled water tune
Zero, measure each pipe absorbance value.
Calculation formula:
VC content (mcg/ml)=(measurement OD value-blank OD value)/(standard OD value-blank OD value) * standard items are dense
Spend (6 mcg/ml) * Sample pretreatment extension rate (4 times) * 4；
3, chlorophyll measuring method measuring principle
Chlorophyll a and chlorophyll b are respectively provided with maximum absorption band in 645nm and 663nm, according to chlorophyll extracting solution pair
The absorption of visible spectrum measures its absorbance in maximum absorption wavelength using spectrophotometer, to calculate chlorophyll a, Ye Lv
Plain b and Chlorophyll content.
Instrument and equipment: visible spectrophotometer, 1ml glass cuvette, 10ml teat glass, mortar, balance, masking foil,
Dehydrated alcohol, acetone and distilled water.
Reagent, pulvis
Operating procedure:
(1) fresh plant blade is taken, middle arteries are removed for plant leaf blade, 0.1g is weighed and shreds, distilled water cleans up.
(2) dehydrated alcohol and acetone are mixed well according to 1:2 (v:v) ratio and step 1 is handled into sample for use for extracting solution
1ml distilled water is added in this, a small amount of pulvis (50mg) is fully ground under the conditions of being protected from light and is placed in 10ml teat glass.
(3) mortar is rinsed with extracting solution, washing lotion is transferred to teat glass, extracting solution is settled to 10ml, is being protected from light condition
Lower extraction about 3h is placed in glass using masking foil if it cannot be protected from light and tries until observing that bottom residues bleach lower extraction completely
Shield lights on pipe.
(4) leaching liquor extracts 1ml and 1ml glass cuvette, and extracting solution zeroing measures extinction at 663 and 645nm respectively
Degree, respectively A663 and A645.
Calculation formula:
Chlorophyll-a Content (mg/g fresh weight)=(12.7 × A663-2.69 × A645) × V mentions (10ml) × extension rate
÷m/(g)÷1000；
Content of chlorophyll b (mg/g fresh weight)=(22.9 × A645-4.68 × A663) × V mentions (10ml) × extension rate
÷m/(g)÷1000；
Chlorophyll content (mg/g fresh weight)=(20.21 × A645+48.02 × A663) × V proposes (10ml) × dilution times
Number ÷ m/ (g) ÷ 1000；
4, method of protein measurement measuring principle
Protein molecule have-NH3+ group, when brownish red Coomassie brilliant blue color developing agent be added protein standard liquid or
When in sample, the anion on Coomassie Brilliant Blue dye is combined with albumen-NH3+, and solution is made to become blue, is inhaled by measurement
Luminosity can calculate protein content.
Reagent and preparation
Reagent: Coomassie brilliant blue stock solution；
The preparation of Coomassie brilliant blue developing solution: in Coomassie brilliant blue stock solution: distilled water=1:4 ratio is prepared,
It prepares on demand, matching while using.
Operating procedure:
(1) Sample pretreatment
Take tissue block (0.2g-0.5g) at least can be to 5-10mg in ice-cold PBS (PH7.4 phosphate buffer)
Rinsing, filter paper are wiped dry, and correct amount is put into the homogenate tube of 5ml.
(2) by weight (g): the homogenate medium of 4 times of volumes is added in homogenate tube in volume (ml)=1:4 ratio, ice water
Under the conditions of bath, tissue block is shredded as early as possible with small cut of ophthalmology.
(3) mode being homogenized: manual homogenization
Left hand holds homogenate tube and fills lower end insertion in the vessel of mixture of ice and water, and stamp stem is inserted perpendicularly into casing by the right hand
In, tens of times of grinding (6-8 minutes) is rotated upwardly and downwardly, sufficiently grinds, 20% homogenate is made.
(4) by the 20% homogenate generic centrifuge prepared or 4000 revs/min of low temperature low speed centrifuge, it is centrifuged 10-
15 minutes, supernatant is taken to be measured.
Operate table:
| Blank tube | Standard pipe | Measurement pipe | |
| Distilled water (ml) | 0.05 | ||
| 0.563g/L protein standard substance (ml) | 0.05 | ||
| Sample (ml) | 0.05 | ||
| Coomassie brilliant blue developing solution (ml) | 3.0 | 3.0 | 3.0 |
It mixes, stands 10 minutes, wavelength 595nm, optical path 1cm, distilled water zeroing measures each pipe absorbance value.
Calculation formula:
Sample to be tested protein concentration (gprot/L)=" (measurement OD value-blank OD value)/(standard OD value-blank OD value) "
Extension rate before × standard concentration (0.563g/L) × sample measures；
5, plant soluble sugar measuring method reagent and preparation
5.1, reagent one: acne；
Reagent two: liquid ' '
The preparation of substrate solution: 5ml reagent two being added in one acne of reagent, sufficiently dissolves, can low-grade fever such as compared with indissoluble solution
It stirs or rocks, stand-by after dissolution, 4 DEG C of the substrate solution prepared is kept in dark place one week.
1mg/ml standard items stock solution 0.3ml；Standard dilutions 10ml
The preparation of 100 mcg/ml standard items application liquid: taking 1mg/ml standard items stock solution 0.2ml, is added
It (is i.e. standard items stock solution: standard dilutions=1: 9), mixes, is prepared into the standard dilutions of 1.8ml
The standard items application liquid of 100 mcg/mls.The concentrated sulfuric acid
5.2, sugar can generate alditol or hydroxymethylfurfural, the alditol or hydroxyl of generation through dehydration under concentrated sulfuric acid effect
Methylfurfural can generate blue-green furfural derivatives with anthrone reaction, in a certain range, the content of the depth of color and sugar at
Direct ratio, therefore can be used for quantifying for sugar.The coloring matter that carbohydrate and anthrone reaction generate is in the absorption peak of visible region
630nm can carry out colorimetric at this wavelength.
5.3, operating procedure
The extraction of soluble sugar: weighing sample to be tested 0.1-0.2g, (fresh plant blade, dries surface contaminants, shreds mixed
It is even), according to sample to be tested quality (g): distilled water is added in the ratio of distilled water (ml)=1: 10, is ground into homogenate with homogenizer
Liquid pours into and changes in centrifuge tube, and boiling water bath 10min (is covered tightly, pipe, which covers, pricks an aperture, with equilibrium air pressure and reduces water dispersion
Lose), after cooling, 4000 revs/min, room temperature is centrifuged 10min, takes 10 times of supernatant distilled water dilutions, shakes up and prepare on sample
Clear liquid is spare.
Operate table
Note: it needs first to do preliminary experiment before formal experiment, if sample to be tested supernatant A value (=A measured value-A blank tube) is greater than
1, it is measured after needing to mix the sample with distilled water dilution, multiplied by extension rate when calculating.
Calculation formula:
Solvable sugared content (micro- gram gram of weight in wet base)=(measurement OD- blank OD)/(standard OD- blank OD) * standard concentration
Extension rate before (100 mcg/ml) ÷ sample to be tested fresh weight/(10 × homogenate distilled water volume ml) × sample measures.
6, result and analysis
The measurement of 6.1 growth potentials, as shown in Figure 3；
6.2 biomass estimations, as shown in Figure 4；
The fresh weight of 8:1 light processing and on the ground weight maximum, but it is not significant (P=0.306) with the difference of 20:5:1 processing,
The significant difference respectively handled with other, (P < 0.05)；20:5:1 processing fresh weight and on the ground take second place again, and with it is red, white (right
According to), blue significant difference (P<0.05), blue light is minimum, but with not significant P>0.05 of white light difference), and it is aobvious with other light processings
Write (P < 0.05) root/shoot ratio trend and fresh weight and weight on the ground it is substantially opposite, control highest, blue light takes second place, between the two with
And difference is significant (P < 0.05) between the two and other light processings, minimum is feux rouges, and with the not significant (P=of 4:1:1 difference
0.245), significant (P=0.054) with the edge 20:5:1, with the equal significant difference (P < 0.05) of other fluorescent tubes processing.
The measurement of 6.3 quality of vegetable
Such as Fig. 5, shown, chlorophyll a, b and Chlorophyll content between the processing of different light without significant difference (P >
0.05)。
As shown in fig. 6, Vc content, which can be seen that the processing of 20:5:1 light, is significantly higher than other processing (P < 0.05), other
It is not significantly different between each processing.
As shown in fig. 6, the processing of protein content 20:5:1 light is significantly higher than complete red, 4:1,4:1:1 and white light control (P
< 0.05), the protein content of full blue light processing is also relatively high, is only second to the processing of 20:5:1 light, and difference is not shown between the two
It writes (P=0.296), but significant (P < 0.05) with Quan Hong, 4:1 and white light contrast difference, and the protein of 8:1 and complete red processing
Content edge is significant (P=0.05), and the protein content of complete red processing is minimum.
As shown in fig. 6, soluble sugar content 20:5:1 highest, but with the difference of complete red and full blue light processing not significantly (P >
0.05), it is significantly higher than other each light processing (P < 0.05), followed by full blue light and the processing of full feux rouges, difference is not between the two
Significantly (P=0.28), but difference is significant between other processing, (P<0.05), between other four kinds processing difference it is not significant (P>
0.05)。
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (9)
1. a kind of ice plant water planting implantation methods, which is characterized in that the ice plant water planting implantation methods include: red
Light: blue light: ultraviolet light=20:5:1 light quality proportion, intensity of illumination are 300 μm of ol/ (m2S), photoperiod 14h/d, Indoor Temperature
Degree is 20-22 DEG C.
2. ice plant water planting implantation methods as described in claim 1, which is characterized in that the ice plant water planting kind
Plant method includes: to carry out nursery using nursery sponge, and sponge is saturating with water-wet, and seed is directly sowed in nursery sponge, is protected from light
Processing 2 days, seed are sprouted, and give illumination later, when seedling it is long to two leaves wholeheartedly when, select the consistent seedling of growing way to carry out water planting
Test；Nutrient solution uses Hogland nutrient solution, solid cultivation, and water planting frame is divided into 3 layers, and 1 layer can plant ice dish 36, every layer of setting
A kind of light source；Since after transplanting 15 days, high every the growth indexes of survey in 10 days, including stem, leaf logarithm, blade was long, blade
Width, hat width is long and hat width is wide, continuous measurement 4 times, and plant is collected to and measured every plant of fresh weight and is planted on the ground, and calculates root cap
Than；Collected material is subjected to Vc, chlorophyll a, b in laboratory, the measurement of total protein and total reducing sugar ingredient, every layer takes 6 samples
Product are measured, and are averaged, 3 repetitions of every kind of light.
3. ice plant water planting implantation methods as claimed in claim 2, which is characterized in that the Vc measuring method includes:
(1) Sample pretreatment takes tissue block 0.2g-0.5g that can at least rinse in ice-cold PBS to 5-10mg, and filter paper is wiped
Dry, weighing is put into the homogenate tube of 5ml；
(2) the homogenate medium of 4 times of volumes is added in homogenate tube in g by weight: volume ml=1:4 ratio, under the conditions of ice-water bath,
Tissue block is shredded as early as possible with small cut of ophthalmology；
(3) mode being homogenized: manual homogenization, left hand hold homogenate tube and fill lower end insertion in the vessel of mixture of ice and water, the right hand
Stamp stem is inserted perpendicularly into casing, is rotated upwardly and downwardly grinding tens of times, is sufficiently ground, 20% homogenate is made；
(4) by the 20% homogenate generic centrifuge prepared or 4000 revs/min of low temperature low speed centrifuge, 10-15 points are centrifuged
Clock takes supernatant to be measured；
(5) prepared by supernatant: take sample 0.15ml reagent adding one to apply liquid 0.45ml, vortex mixes, and it is centrifuged after placing 15 minutes,
It 3500-4000 revs/min, is centrifuged 10 minutes, upper layer clear liquid is supernatant.
4. ice plant water planting implantation methods as claimed in claim 3, which is characterized in that the survey of VC in the supernatant
Fixed: 37 DEG C mix well for water-bath 30 minutes, stand 10 minutes, wavelength 536nm, optical path 1cm, and distilled water zeroing measures each pipe and inhales
Shading value；
Calculation formula:
VC content (mcg/ml)=(measurement OD value-blank OD value)/(standard OD value-blank OD value) (6 is micro- for * standard concentration
Grams per milliliter) * Sample pretreatment extension rate (4 times) * 4.
5. ice plant water planting implantation methods as claimed in claim 2, which is characterized in that the chlorophyll measuring method packet
It includes:
(1) fresh plant blade is taken, middle arteries are removed for plant leaf blade, 0.1g is weighed and shreds, distilled water cleans up；
(2) 1ml will be added in processing sample for use for extracting solution by mixing well dehydrated alcohol and acetone according to 1:2v:v ratio
Distilled water, pulvis 50mg are fully ground under the conditions of being protected from light and are placed in 10ml teat glass；
(3) mortar is rinsed with extracting solution, washing lotion is transferred to teat glass, extracting solution is settled to 10ml, soaks under the conditions of being protected from light
3h is mentioned, until observing that bottom residues bleach lower extraction completely, is placed on teat glass and is hidden using masking foil if it cannot be protected from light
Opacus line；
(4) leaching liquor extracts 1ml and 1ml glass cuvette, and extracting solution zeroing measures absorbance at 663 and 645nm respectively, point
It Wei not A663 and A645；
Calculation formula:
Chlorophyll-a Content (mg/g fresh weight)=(12.7 × A663-2.69 × A645) × V mentions (10ml) × extension rate ÷ m/
(g)÷1000；
Content of chlorophyll b (mg/g fresh weight)=(22.9 × A645-4.68 × A663) × V mentions (10ml) × extension rate ÷ m/
(g)÷1000；
Chlorophyll content (mg/g fresh weight)=(20.21 × A645+48.02 × A663) × V mentions (10ml) × extension rate ÷
m/(g)÷1000。
6. ice plant water planting implantation methods as claimed in claim 2, which is characterized in that the method for protein measurement packet
It includes:
(1) Sample pretreatment takes tissue 0.2g-0.5g) can at least be rinsed in ice-cold PBS to 5-10mg, filter paper wipe it is dry,
Correct amount is put into the homogenate tube of 5ml；
(2) the homogenate medium of 4 times of volumes is added in homogenate tube in g by weight: volume ml=1:4 ratio, under the conditions of ice-water bath,
Tissue block is shredded as early as possible with small cut of ophthalmology；
(3) mode being homogenized: manual homogenization, left hand hold homogenate tube and fill lower end insertion in the vessel of mixture of ice and water, the right hand
Stamp stem is inserted perpendicularly into casing, is rotated upwardly and downwardly grinding tens of times, is sufficiently ground, 20% homogenate is made；
(4) by the 20% homogenate generic centrifuge prepared or 4000 revs/min of low temperature low speed centrifuge, 10-15 points are centrifuged
Clock takes supernatant to be measured.
7. ice plant water planting implantation methods as claimed in claim 6, which is characterized in that the measuring method of the supernatant
Include: to mix, stand 10 minutes, wavelength 595nm, optical path 1cm, distilled water zeroing measures each pipe absorbance value；
Calculation formula:
Sample to be tested protein concentration (gprot/L)=" (measurement OD value-blank OD value)/(standard OD value-blank OD value) " × mark
Extension rate before quasi- product concentration (0.563g/L) × sample measures.
8. ice plant water planting implantation methods as claimed in claim 2, which is characterized in that the plant soluble sugar measurement
Method includes: to weigh sample to be tested 0.1-0.2g, and distillation is added according to sample to be tested quality g: distilled water ml=1:10 ratio
Water is ground into homogenate with homogenizer, pours into and changes in centrifuge tube, boiling water bath 10min；It covers tightly, pipe, which covers, pricks an aperture, cooling
Afterwards, 4000 revs/min, room temperature is centrifuged 10min, takes 10 times of supernatant distilled water dilutions, shakes up that prepare sample supernatant spare；
Calculation formula:
Solvable sugared content (micro- gram gram of weight in wet base)=(measurement OD- blank OD)/(standard OD- blank OD) (100 is micro- for * standard concentration
Grams per milliliter) ÷ sample to be tested fresh weight/(10 × it is homogenized with extension rate before distilled water volume ml) × sample measures.
9. a kind of measuring method of the LED light matter proportion of ice plant water planting plantation as described in claim 1, feature exist
In, the LED light matter proportion of ice plant water planting plantation measuring method the following steps are included:
Step 1, experiment carry out 3 repetitions, repeat that 7 kinds of light qualities are arranged every time, feux rouges, blue light, feux rouges: blue light=4:1, feux rouges:
Blue light=8:1, feux rouges: blue light: green light=4:1:1, feux rouges: blue light: ultraviolet light=20:5:1, white light control；
Step 2, ice dish growth potential and growth curve under handling different light medium proportion are measured；
Step 3 weighs to the biomass of the ice dish under different light medium proportion processing；
Step 4 handles different light medium proportion the chlorophyll a of lower ice dish, and chlorophyll b, Chlorophyll content, vitamin C can
The content of dissolubility albumen and soluble sugar is measured.
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